Characterization of unique truncated prolactin receptor transcripts, corresponding to the intracellular domain, in the testis of the sexually mature chicken.
Characterization of unique truncated prolactin receptor transcripts, corresponding to the intracellular domain, in the testis of the sexually mature chicken.
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DOI：
10.1210/endo.140.3.6603
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发表时间：
1999-03
期刊：
影响因子：
4.8
通讯作者：
J. Mao;J. Burnside;L. Li;J. Tang;C. Davolos;L. Cogburn
中科院分区：
文献类型：
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作者：
J. Mao;J. Burnside;L. Li;J. Tang;C. Davolos;L. Cogburn
我们用北方杂交技术检测了鸡催乳素受体（cPRLR）基因在鸡不同组织中的表达。检查的大多数组织（卵巢、睾丸、输卵管、肾脏和脂肪）具有显著的全长（4.6 kb）cPRLR转录物。一个更大的（11.7 kb）的成绩单也检测到卵巢，输卵管，睾丸和肾脏后，更长的曝光。在性成熟鸡的睾丸中发现了cPRLR表达的独特模式，除了在其他组织中发现的4.6 kb转录物外，睾丸中还具有异常高的三个小转录物（1.2，1.7和2 kb）丰度。构建了三个结构域特异性互补DNA（cDNA）探针，其对应于胞外结构域和跨膜-胞内结构域中的第一和第二配体结合区。用这些探针，北方印迹分析从成熟（22周龄）鸡睾丸制备的多聚腺苷酸化RNA表明，睾丸中高丰度（1.2-和1.7-kb）和低丰度（2.0-kb）cPRLR转录物仅与细胞内结构域探针杂交。采用5 ′-RACE（rapid amplification of cDNA ends）技术对22周龄鸡睾丸组织中的多聚腺苷酸化RNA进行分析，鉴定出两种睾丸特异性截短型cPRLR转录本。主要的截短cDNA序列含有高度保守的box 1基序[（+）box 1 cDNA]，并与cPRLR cDNA的序列（位于跨膜结构域下游）发生分歧（在核苷酸1396处）。另一个截短的cDNA缺少box 1基序[（-）box 1cDNA]，其被39个碱基取代，可以编码具有131个碱基的推定的5 '非翻译区的疏水性N-末端。幼龄鸡主要在睾丸中表达全长cPRLR信使RNA（4.6 kb）。在性成熟开始时，睾丸特异性（+）box 1转录物的丰度急剧增加，而全长cPRLR的表达受到抑制。在性成熟的鸡睾丸中存在截短的[（+）或（-）box 1] cPRLR转录本，表明PRL对性腺功能的作用机制复杂。
We have examined expression of the chicken PRL receptor (cPRLR) gene in different tissues of the chicken by Northern blot analysis. Most tissues examined (ovary, testis, oviduct, kidney, and fat) possess a prominent full-length (4.6-kb) cPRLR transcript. A larger (11.7-kb) transcript is also detected in ovary, oviduct, testis, and kidney after longer exposure. A unique pattern of cPRLR expression was found in the testis of sexually mature chickens, which have an unusually high abundance of three small transcripts (1.2, 1.7, and 2 kb) in addition to the 4.6-kb transcript found in other tissues. Three domain-specific complementary DNA (cDNA) probes were constructed that correspond to the first and second ligand-binding regions in the extracellular domain and the transmembrane-intracellular domain. With these probes, Northern blot analysis of polyadenylated RNA prepared from the testes of a mature (22-week-old) chicken indicates that the highly abundant (1.2- and 1.7-kb) and less abundant (2.0-kb) cPRLR transcripts in testis hybridize only to the intracellular domain probe. Two types of truncated testis-specific cPRLR transcripts were identified using 5'-RACE (rapid amplification of cDNA ends) analysis of polyadenylated RNA from the testis of a 22-week-old chicken. The predominant truncated cDNA sequence contains the highly conserved box 1 motif [(+)box 1 cDNA] and diverges (at nucleotide 1396) from that of the cPRLR cDNA, just downstream of the transmembrane domain. The other truncated cDNA lacks the box 1 motif [(-)box 1 cDNA], which is replaced by 39 bases that could encode a hydrophobic N-terminus with a putative 5'-untranslated region of 131 bases. Young chickens predominately express the full-length cPRLR messenger RNA (4.6 kb) in the testis. At the onset of sexual maturity, there is a dramatic increase in abundance of the testis-specific (+)box 1 transcript, whereas expression of the full-length cPRLR is depressed. The presence of truncated [(+) or (-)box 1] cPRLR transcripts in the sexually mature chicken testis suggests a complex mechanism of PRL action on gonadal function.